CN201418905Y

CN201418905Y - Fiber mesh stacked bioreactor for artificial liver

Info

Publication number: CN201418905Y
Application number: CN2009201193491U
Authority: CN
Inventors: 李兰娟; 朱春侠; 喻成波; 曹红翠; 杜维波; 潘小平
Original assignee: Zhejiang University ZJU
Current assignee: Zhejiang University ZJU
Priority date: 2009-05-07
Filing date: 2009-05-07
Publication date: 2010-03-10
Anticipated expiration: 2019-05-07

Abstract

The utility model relates to medical equipment, aiming to provide a fiber mesh stacked bioreactor for an artificial liver. The reactor includes a reactor shell, in which several vertically stacked hollow fiber mesh sheets are arranged, the hollow fiber mesh sheets are woven by hollow fibers, and several vertically arranged hollow fiber mesh sheets pass through the grid of the aforementioned hollow fiber mesh sheets The hollow fibers are all polysulfone hollow fibers; the bottom of the reactor shell is provided with a cell suspension inlet and a liquid inlet, the top is provided with a cell suspension outlet and a liquid outlet, and the side walls are oppositely provided with a gas inlet and a gas outlet. The utility model has a three-dimensional culture structure, has oxygenation and immune barrier functions, has good cell activity and function, and is easy to enlarge.

Description

Artificial liver fiber mat superposed type biological reactor

Technical field

This utility model relates to a kind of armarium, specifically, is the core apparatus of a kind of biotype or hybrid artificial liver treatment, i.e. artificial liver fiber mat superposed type biological reactor.

Background technology

Bioreactor (being reactor) is the core apparatus of biotype and hybrid artificial liver treatment, and the place of mass exchange both is provided for exogenous hepatocyte and blood samples of patients or blood plasma, for hepatocyte provides suitable growing environment, has a extensive future again.Ideal bioreactor for artificial liver should meet following basic demand: 1, competent hepatic cell growth space, and support a liver failure patient to need 10% normal liver quality approximately, amount to hepatocyte number about 15,000,000,000.2, excellent biological compatibility can guarantee hepatocellular activity and function.3, medium and small material transmitted in both directions, the macromolecular substances immunity intercepts.

Bioreactor mainly contains four types so far: hollow fiber type, dull and stereotyped monolayer culture type, filling fluidized-bed type/support rack type, cell parcel/floating type.The hollow fiber type reactor is convenient to the hepatocyte adherent growth because have the surface of sticking, and immune iris action is arranged, and cell is subjected to advantages such as shearing force is little, is to use maximum types at present.But there is following shortcoming in it: 1, in the traditional hollow fiber reactor, no matter the parallel tight arrangement of fiber is to cultivate outside intracavity or chamber, and the hepatic cell growth space is all inadequate; 2, can only cultivate for hepatocyte provides two dimension, can not provide 3 D stereo to cultivate, cytoactive and function are not good, are difficult at systematism; 3, cell distribution inequality; 4, cell easily stops up semipermeable membrane, influences mass exchange.Irritate fluidized-bed type/support rack type reactor, have and can amplify easily for hepatocyte provides the dimensional culture environment, the advantage that mass exchange is good is used also morely aspect zoopery and cell culture, but has following problem: 1, lack immune isolated potential safety hazard; 2, mostly the internal stent hole is irregular uneven, easily produces the perfusion inequality, and the center of support integral body and edge zone easily produce dead space, dead space; 3, because big by the resistance of a large amount of supports, required perfusion intensity is big, causes cell and is subjected to shearing force big, is difficult to amplify; 4, the making of reactor is subjected to the restriction of timbering material processing technology.And dull and stereotyped monolayer culture type reactor since body surface area than little, amplify complicatedly, cell parcel floating type is because the stability of suspension cell is not good, the mass exchange of parcel cell is influenced, the research report is less recently.

How to develop and improve bioreactor, maximize favourable factors and minimize unfavourable ones, overcome one's shortcomings, become recently the focus of research both at home and abroad.

The utility model content

The purpose of this utility model is to provide a kind of to be possessed 3 D stereo and cultivates structure, has oxygenate concurrently and immune barriers function, cytoactive and function artificial liver fiber mat superposed type biological reactor good, that be easy to amplify.

Artificial liver of the present utility model fiber mat superposed type biological reactor, comprise shell of reactor, arrange some vertical synergetic doughnut net sheets in the shell of reactor, doughnut net sheet is formed by woven hollow fiber, the doughnut of some vertical layouts passes the center of the grid of aforementioned doughnut net sheet, and described doughnut is polysulfone hollow fibre; Cell suspension inlet and liquid inlet are established in the bottom of shell of reactor, and cell suspension outlet and liquid outlet are established in the top, and the sidewall subtend is provided with gas access and gas outlet;

The described inside that is used to weave the doughnut of doughnut net sheet is gas passage, and the one end connects the gas access, and the other end connects gas outlet; The inside of the doughnut of described vertical layout is fluid passage, and the one end connects the liquid inlet, and the other end connects liquid outlet; The hepatocellular three dimensional growth of the common formation of the outer wall of all doughnuts and the inwall of shell of reactor space is communicated with cell suspension outlet and cell suspension inlet.

Bacteria filter all is equipped with in described cell suspension inlet and cell suspension exit, and is fixed on the shell of reactor with spiral cover.

Polysulfone hollow fibre or polyether sulfone doughnut that the fibronectin that the described doughnut that is used to weave doughnut net sheet is a permeable watertight is modified.

The doughnut of described vertical layout is aperture 0.2 μ m, molecular retention amount 50～100kD and not modified polysulfone hollow fibre or polyether sulfone doughnut.

The grid length of side of described doughnut net sheet is 1mm.

Described vertical synergetic doughnut net sheet is to be formed by stacking according to the spacing of 300 μ m is bonding.

Described shell of reactor is cuboid or cylindrical transparent lucite shell.

The gas access and the gas outlet of the setting of described shell of reactor sidewall subtend have two respectively, and two groups of gas accesses and gas outlet be the doughnut that is used to weave doughnut net sheet of a corresponding direction respectively, constitutes independently gas passage.

The beneficial effect that the utlity model has is:

Utilize the spatial organization of doughnut, do not introduce three-dimensional biological support, but realized the 3 D stereo cultivation, help the active raising of hepatocyte function and keep for a long time, be easy to systematism; The three-dimensional hole rule that fiber combinations constitutes is even, and liquid flowing resistance is little, and the suffered shearing force of cell is little, and cell distribution is more even; Utilize the combination of different qualities doughnut, make oxygenate fully, have immune barriers function concurrently, mass exchange efficient height has improved the defective that cell easily stops up semipermeable membrane.Simple and compact for structure, used doughnut material technical maturity, practical, be easy to amplify.

Description of drawings

Fig. 1 is a reactor longitudinal profile structural representation of the present utility model;

Fig. 2 is a reactor longitudinal profile structural representation of the present utility model;

Fig. 3 uses sketch map for the biotype artificial liver support system.

Reference numeral: doughnut net sheet 1, the vertical doughnut of arranging 2, hepatocellular three dimensional growth space 3, shell of reactor 4, liquid inlet 5, liquid outlet 6, gas access 7, gas outlet 8, cell suspension inlet 9, cell suspension outlet 10, bacteria filter 11, spiral cover 12, patient or laboratory animal 13, plasma separator 14, extracorporeal circulation pool 15.

The specific embodiment

Artificial liver of the present utility model fiber mat superposed type biological reactor, the spacing of doughnut net sheet, sizing grid and reactor scale can or be dwindled according to the practical application amplification.

Below in conjunction with the specific embodiment this utility model is described in further detail:

Artificial liver in present embodiment fiber mat superposed type biological reactor comprises the shell of reactor 4 of a transparent organic glass.Cell suspension inlet 9 and liquid inlet 5 are established in the bottom of shell of reactor 4, and cell suspension outlet 10 and liquid outlet 6 are established in the top, and the sidewall subtend is provided with gas access 7 and gas outlet 8.Cell suspension inlet 9 and cell suspension export 10 places bacteria filter 11 all are housed, and are fixed on the shell of reactor 4 with spiral cover 12.

Arrange some vertical synergetic doughnut net sheets 1 in the shell of reactor 4, doughnut net sheet 1 is formed by the polysulfone hollow fibre braiding that the surface is fit to fibronectin modification hepatic cell growth, permeable watertight.The grid length of side of gathering on the net sheet is 1mm, and each layer net sheet is formed by stacking according to the spacing of 300 μ m is bonding.The inside of the doughnut of doughnut net sheet 1 is the passage of oxygen and carbon dioxide gas mixture, and the one end connects gas access 7, and the other end connects gas outlet 8.

The gas access 7 and the gas outlet 8 of the subtend setting of shell of reactor sidewall middle part have two respectively, and two groups of gas accesses 7 and gas outlet 8 be the doughnut that is used to weave doughnut net sheet 1 of a corresponding direction respectively, constitutes independently gas passage.Can certainly on chamber wall cuts off, design, only adopt one group of gas access 7 and gas outlet 8.

The doughnut 2 of some vertical layouts passes the center of the grid of doughnut net sheet 1, the vertical doughnut of arranging 2 is aperture 0.2 μ m, molecular retention amount 50～100kD and not modified polysulfone hollow fibre, possess the good and immune isolation effect of biocompatibility, and can reduce hepatocyte is stopped up fiber holes in the quick growth in its surface risk.The inside of the vertical doughnut of arranging 2 is passages of blood plasma or culture fluid, and the one end connects the liquid inlet, and the other end connects liquid outlet.

The characteristics of the doughnut of doughnut net sheet 1 are: permeable watertight, and the surface is easy to hepatic cell growth; The characteristics of the vertical doughnut of arranging 2 are: air-and water-permeable, have immune iris action, have good biocompatibility, the surface needn't be easy to hepatic cell growth very much, and is too fast and stop up the risk of fiber holes in its superficial growth to reduce cell.According to this standard, also can select for use polyether sulfone etc. to possess the material of same characteristic as doughnut.

The hepatocellular three dimensional growth of the common formation of the inwall of the outer wall of all doughnuts and shell of reactor 4 space 3, this space is communicated with cell suspension inlet 9 and cell suspension outlet 10.Hepatocyte mainly attaches to the surface of doughnut net sheet 1, and part attaches to the surface of the doughnut 2 of vertical layout, is the 3 D stereo growth, is easy to systematism.

Interlacing part doughnut in reactor can remove with cell suspension inlet 9 and cell suspension outlet 10, so that the inflow of hepatocyte suspension and outflow.

Use in the present embodiment is as follows:

After the blood lead body of patient or laboratory animal 13 is outer, at first be separated into cell composition and blood plasma components through blood plasma separator 14, wherein blood plasma components enters extracorporeal circulation pool 15,5 enter in the reactor from the liquid inlet then, inner chamber by vertical doughnut 2 flows out, after hepatocyte in the hepatocellular three dimensional growth space 3 (hepatocyte for inoculate in advance and cultivate) fully contacts, flow back to the inner chamber of vertical doughnut 2, liquid outlet 6 to reactor head flows out from bottom to top, returns circulatory pool 15.After repeatedly circulating, the cell composition in blood feeds back in the body again.

Hepatocyte can be selected former generation people, porcine hepatocyte or hepatic cell line etc. for use, and summary inoculation and incubation step are as follows:

1, is mixed with the hepatocyte suspension of suitable concn with culture fluid.

2, open the spiral cover 12 of cell suspension inlet 9, in the hepatocyte suspension injecting reactor, till being full of reactor and overflowing to cell suspension from the cell suspension outlet 10 at top.Hepatocyte attaches to the doughnut outer wall, attaches firmly after about 24 hours.

3, pour into fresh culture from cell suspension inlet 9, flow out from cell suspension outlet 10, wash away not adherent hepatocyte through reactor.

4, shown in pressing this reactor and extracorporeal circulation pool 15 etc. in Fig. 3 frame of broken lines, be linked to be the liver cell culture peripheral passage, fresh culture medium respectively from the liquid inlet 5 and liquid outlet 6 flow into and outflow reactors, for hepatocyte provides nutrient and takes away metabolite.

5, after the suitable time of cultivation, can be combined into the Biotype artificial rami hepatici by Fig. 3 connection and hold system.

Claims

1. A fiber mesh superimposed bioreactor for an artificial liver, comprising a reactor shell, characterized in that a number of vertically stacked hollow fiber mesh sheets are arranged in the reactor casing, and the hollow fiber mesh sheets are woven by hollow fibers. Several vertically arranged hollow fibers pass through the center of the grid of the aforementioned hollow fiber mesh, and the hollow fibers are polysulfone hollow fibers or polyethersulfone hollow fibers; the bottom end of the reactor shell is provided with a cell suspension inlet and a liquid inlet , the top is provided with a cell suspension outlet and a liquid outlet, and the side walls are oppositely provided with a gas inlet and a gas outlet;

The inside of the hollow fiber used to weave the hollow fiber mesh is a gas channel, one end of which is connected to the gas inlet, and the other end is connected to the gas outlet; the inside of the vertically arranged hollow fiber is a liquid channel, one end of which is connected to the liquid inlet, The other end is connected to the liquid outlet; the outer walls of all the hollow fibers and the inner wall of the reactor shell jointly form a three-dimensional growth space for liver cells, which communicates with the cell suspension outlet and the cell suspension inlet.

2. The fiber mesh superimposed bioreactor for artificial liver according to claim 1, characterized in that, the cell suspension inlet and the cell suspension outlet are equipped with bacteria filters, which are fixed to the reactor with screw caps on the casing.

3. The fiber mesh superimposed bioreactor for artificial liver according to claim 1, characterized in that, the hollow fibers used to weave the hollow fiber mesh are air-permeable and impermeable fibronectin-modified polysulfone hollow fiber or polyethersulfone hollow fiber.

4. The fiber mesh stacked bioreactor for artificial liver according to claim 1, wherein the vertically arranged hollow fibers are unmodified polymers with a pore size of 0.2 μm and a molecular cut-off of 50-100 kD. Sulfone hollow fibers or polyethersulfone hollow fibers.

5. The fiber mesh superimposed bioreactor for artificial liver according to claim 1, characterized in that the mesh side length of the hollow fiber mesh is 1 mm.

6. The fiber mesh superimposed bioreactor for artificial liver according to claim 1, characterized in that, the vertically stacked hollow fiber mesh is bonded and stacked at a distance of 300 μm.

7. The fiber mesh stacked bioreactor for artificial liver according to claim 1, wherein the shell of the reactor is a cuboid or a cylinder.

8. The fiber mesh superimposed bioreactor for artificial liver according to claim 1, characterized in that the shell of the reactor is a transparent plexiglass shell.

9. The fiber mesh stacked bioreactor for artificial liver according to claim 1, characterized in that there are two gas inlets and two gas outlets opposite to each other on the side wall of the reactor shell, and two sets of gas inlets The hollow fiber used to weave the hollow fiber mesh sheet corresponding to one direction respectively with the gas outlet constitutes an independent gas channel.